Saxitoxins induce cytotoxicity, genotoxicity and oxidative stress in teleost neurons in vitro

被引：26

作者：

da Silva, Cesar Aparecido ^{[1
]}

Penna de Morais, Elizabeth Cunha ^{[2
]}

Moura Costa, Michele Dietrich ^{[2
]}

Coelho Ribas, Joao Luiz ^{[3
]}

Guiloski, Izonete Cristina ^{[3
]}

Ramsdorf, Wanessa A. ^{[4
]}

Zanata, Silvio Marques ^{[2
]}

Cestari, Marta M. ^{[4
]}

Oliveira Ribeiro, Ciro Alberto ^{[5
]}

Magalhaes, Valeria E. ^{[6
]}

Trudeau, Vance L. ^{[7
]}

Silva de Assis, Helena C. ^{[3
]}

机构：

[1] Univ Fed Parana, Ecol & Conservat Postgrad Program, BR-81531990 Curitiba, Parana, Brazil

[2] Univ Fed Parana, Dept Basic Pathol, BR-81531990 Curitiba, Parana, Brazil

[3] Univ Fed Parana, Dept Farmacol, BR-81531990 Curitiba, PR, Brazil

[4] Univ Fed Parana, Dept Genet, BR-81531990 Curitiba, Parana, Brazil

[5] Univ Fed Parana, Dept Cellular Biol, BR-81531990 Curitiba, Parana, Brazil

[6] Univ Fed Rio de Janeiro, Inst Biophys Carlos Chagas Filho, BR-21941902 Rio De Janeiro, RJ, Brazil

[7] Univ Ottawa, Dept Biol, Ottawa, ON K1N 6N5, Canada

来源：

TOXICON | 2014年 / 86卷

关键词：

Neuron primary culture; Environmental neuroscience; Neurotoxicology; Teleost; FISH HOPLIAS-MALABARICUS; DNA-DAMAGE; METHYL MERCURY; HEPATOCYTES; RESPONSES; CULTURE; CELLS; ASSAY; DDT;

D O I：

10.1016/j.toxicon.2014.04.016

中图分类号：

R9 [药学];

学科分类号：

1007 ;

摘要：

The aim of this study was establish a protocol for isolation and primary culture of neurons from tropical freshwater fish species Hoplias malabaricus for assessment of the effects of neurotoxic substances as saxitoxins (STXs). Cells from brain of H. malabaricus were treated with different concentrations of trypsin, dispase and papain for tissue dissociation. Cells type was separated by cellular gradient and basic fibroblast growth factor (bFGF) supplement nutrition media were added. The dissociated cells were plated with medium and different STXs concentrations and the toxic cellular effects such as oxidative stress, neurotoxicity, and genotoxicity and apoptosis process were evaluated. Cultures treated with bFGF showed the greatest adherence, survival and cellular development. STXs increased specific activity of glutathione peroxidase and lipoperoxidation levels, were cytotoxic and genotoxic indicated by the comet assay. Although the STXs effects due the blockage of sodium channels is reported to be reversible, the time exposure and concentration of STXs suggested cellular injuries which can lead to neuropathology. The establishment of primary neuronal culture protocol enables new applications for neuro-toxicological assessments. (C) 2014 Elsevier Ltd. All rights reserved.

引用

页码：8 / 15

页数：8

共 48 条

[1]

BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3

[2] Isolation and culture of adult neurons and neurospheres [J].

Brewer, Gregory J. ;

Torricelli, John R. .

NATURE PROTOCOLS, 2007, 2 (06) :1490-1498

[3] Responses of hepatocytes to DDT and methyl mercury exposure [J].

Bussolaro, D. ;

Filipak Neto, F. ;

Oliveira Ribeiro, C. A. .

TOXICOLOGY IN VITRO, 2010, 24 (06) :1491-1497

[4] Genetic damage induced by trophic doses of lead in the neotropical fish Hoplias malabaricus (Characiformes, Erythrinidae) as revealed by the comet assay and chromosomal aberrations [J].

Cestari, MM ;

Lemos, PMM ;

Ribeiro, CAD ;

Costa, JRMA ;

Pelletier, E ;

Ferraro, MVM ;

Mantovani, MS ;

Fenocchio, AS .

GENETICS AND MOLECULAR BIOLOGY, 2004, 27 (02) :270-274

[5] Analyses of paralytic shellfish toxins and biomarkers in a southern Brazilian reservoir [J].

Clemente, Zaira ;

Busato, Raquel H. ;

Oliveira Ribeiro, Ciro A. ;

Cestari, Marta M. ;

Ramsdorf, Wanessa A. ;

Magalhaes, Valeria F. ;

Wosiack, Ana C. ;

Silva de Assis, Helena C. .

TOXICON, 2010, 55 (2-3) :396-406

[6] Caspases: the executioners of apoptosis [J].

Cohen, GM .

BIOCHEMICAL JOURNAL, 1997, 326 :1-16

[7]

Collins A, 1997, ENVIRON MOL MUTAGEN, V30, P139, DOI 10.1002/(SICI)1098-2280(1997)30:2<139::AID-EM6>3.0.CO

[8]

2-I

[9] THE INHIBITION OF ISLET SUPEROXIDE-DISMUTASE BY DIABETOGENIC DRUGS [J].

CROUCH, RK ;

GANDY, SE ;

KIMSEY, G ;

GALBRAITH, RA ;

GALBRAITH, GMP ;

BUSE, MG .

DIABETES, 1981, 30 (03) :235-241

[10] First report about saxitoxins in freshwater fish Hoplias malabaricus through trophic exposure [J].

da Silva, Cesar A. ;

Oba, Eliane T. ;

Ramsdorf, Wanessa A. ;

Magalhaes, Valeria E. ;

Cestari, Marta M. ;

Oliveira Ribeiro, Ciro Alberto ;

Silva de Assis, Helena C. .

TOXICON, 2011, 57 (01) :141-147

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